AU2003299772C1 - Association between a PPAR ligand and an antioxidant agent and use thereof for treating obesity - Google Patents

Description

- 1 ASSOCIATION BETWEEN A LIGAND OF PEROXISOME PROLIFERATOR ACTIVATED RECEPTORS AND AN ANTIOXIDANT AGENT, AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM The present invention relates to the association between one or more selective ligands of 5 peroxisome proliferator activated receptors (PPAR) and an antioxidant agent and to the use thereof in the treatment and/or prevention of obesity and of overweight characterised by a body mass index greater than 25. Obesity is a major public health problem in all developed countries. It is also increasing steadily in developing countries and is affecting an ever younger population. Obesity is a 10 chronic disorder of energy imbalance characterised by an excess of energy intake in the long term compared with limited energy expenditure, leading to storage of the excess energy in the form of white adipose tissue. Excess adipose tissue directly contributes to problems of fatigue, shortness of breath, sleep apnoea and osteoarthritis. 15 Obesity is a well-established risk factor for the development of insulin resistance, of dyslipidaemia and, ultimately, of non-insulin-dependent diabetes. It is a factor contributing to cardiovascular diseases and is associated with a significantly increased risk of cerebro vascular accidents, vesicular calculi, respiratory dysfunction, osteoarthritis, several forms of cancer and premature death. 20 A pharmacological strategy for reducing obesity presents two alternatives: either to reduce fat by modifying energy intake and/or by modifying the distribution of nutrients between fat and lean tissues, or to counter or reverse the metabolic consequences of the increase in fat without necessarily having an impact on the degree of obesity in itself. It has been found that, in obese people, the generation of reactive oxygenated species 25 released by monocytes and leukocytes is greatly increased with respect to non-obese -2 subjects (J. Clin. Endocrinol. Metab., 2001, 86, 355-362). Elevated plasma concentrations of alpha tumour necrosis factor (TNFa) in obese people stimulate inflammatory processes (J. Clin. Endocrinol. Metab., 1998, 8, 2907-2910) and are responsible for the generation of reactive oxygenated species by leukocytes (Oncogene, 1998, 17, 1639-1651). 5 The pathological state of obesity is also associated with increased oxidation of lipids and proteins, which may be the cause of high plasma levels of 9- and 13-hydroxy octadecadienoic acids (9-HODE and 13-HODE) (Totowa : Humano. Press., 1998, 147 155), key indices of lipid peroxidation (J. Clin. Endocrinol. Metab., 2001, 86, 355-362). In parallel, the "antioxidant" capabilities of the body are reduced. 10 In obese subjects, it has been shown that excessive food intake causes major lipid and protein damage. Over-consumption of calories by obese people can cause the formation of free radicals and expose them to significant oxidative lesions which help to maintain the state of obesity. The specific markers of oxidation are significantly reduced by a 48-hour fast or by calorie 15 restriction accompanying weight loss (J. Clin. Endocrinol. Metab., 2001, 86, 355-362). A strategy aimed at reducing the "oxidative burden" on the body by favouring the lipid and carbohydrate metabolisms should result in an exacerbation of the effects and, as a consequence, in weight loss in obese or overweight subjects. Among the compounds capable of favouring the lipid and carbohydrate metabolisms, 20 selective ligands of peroxisome proliferator activated receptors or PPARs are especially interesting compounds. The PPARs are a family of nuclear hormone receptors comprising three distinct sub-types: a, P (also called 8 or NUCl) and y (which has three isoforms : y', 72 and 73). They were initially cloned as nuclear receptors mediating the effects of peroxisome 25 proliferators on gene transcription, but it is clear that a very large number of natural compounds such as eicosanoids and fatty acids can also activate PPARs.

-3 Like a certain number of other nuclear hormone receptors, PPAR proteins bind to promoters in the form of heterodimers with the 9-cis-retinoic acid receptor, RXR. The PPAR/RXR heterodimer can be activated by the binding of a ligand specific to one of the two receptors but maximum activation is achieved when two ligands are present. 5 PPARs are ligand-dependent transcription factors, which means that inititation of transcription of the target genes is strictly dependent on the binding of the ligand. Certain ligands, such as mono- or poly-unsaturated fatty acids or saturated fatty acids, bind to the three sub-types of receptor. Long-chain polyunsaturated fatty acids, such as linolenic acid, or oxidated or conjugated fatty acids bind to PPARa with a high degree of affinity. 10 The most important function of PPARs results from their tissue-dependent expression and from their specific target genes which are very often involved exclusively in the transport and metabolism of lipids and carbohydrates. The PPARct KO mouse develops obesity and hypertriglyceridaemia even if the daily intake of calories is not increased. These effects are largely explained by a reduction in 15 fatty acid uptake by the liver and inhibition of fatty acid oxidation (J. Biol. Chem., 1998, 273, 29577-29585). The liver is an organ capable of oxidising fatty acids. When hepatic oxidation of fatty acids is optimal, thermogenesis comes into play and converts the available energy into heat, with a reduction in the respiratory quotient and an increase in the basic metabolic rate. These 20 circumstances are highly favourable to the loss of adipose tissue (Med. Hypotheses, 1999, 52(5), 407-416). A strategy consisting of disinhibiting the enzymatic processes of hepatic oxidation of fatty acids whilst ensuring transcriptional stimulation of genes activated by PPARs and involved in lipid and carbohydrate metabolic processes should result in a reduction in free fatty 25 acids in the plasma and in moderated lipolysis in adipocytes constituting visceral adipose 4 tissue, in the long term bringing about a regression in visceral obesity and, accordingly, a reduction in body weight. The present invention relates, more specifically, to the association between one or more compounds that are ligands of peroxisome proliferator activated 5 receptors and an antioxidant agent. This association exhibits pharmacological properties that are entirely remarkable in the area of obesity. There is disclosed herein a composition comprising 1) an antioxidant agent and 2) a mixed PPAR ligand for the ax and y receptor sub-types, or a selective PPAR 10 ligand for the ax receptor sub-type and a selective PPAR ligand for the y receptor sub-type optionally as a pharmaceutical composition being in combination with one or more pharmaceutically acceptable excipients. More specifically, the PPAR ligands according to the invention are selective ligands for the ax and/or y receptor sub-types. 15 Advantageously, the association according to the invention comprises a selective PPARx ligand and a selective PPARy ligand. An advantageous embodiment relates to the association according to the invention wherein the PPAR ligand is a mixed ligand of the a and y receptor sub types. 20 PPARax and/or y ligands according to the invention are advantageously represented by gemfibrozil, WY-14,643, pioglitazone and, even more preferably, by rosiglitazone. PPAR ligands according to the invention are also represented by the compounds described in the Applications WO 9736579, WO 9731907, W09728115, 5 W09638415, W09727857, W09725042, W09701420, W09640128, W02000064888 and W02000064876. Antioxidant agents according to the invention are represented by various categories of compound : 5 - anti-free radical agents or free-radical trapping agents, - antilipoperoxidant agents, - chelating agents, - agents capable of regenerating endogenous antioxidants such as glutathione, vitamin C or vitamin E. 10 The antioxidant agent of the association according to the invention is more preferably represented by quinone compounds such as ubiquinone or coenzyme Q10, which acts as a free-radical trapping agent but which is also capable of regenerating vitamin E. 15 Also disclosed is a composition, which is rosiglitazone and coenzyme Q 10 optionally as a pharmaceutical composition being in combination with one or more pharmaceutically acceptable excipients. The enantiomers and diastereoisomers and addition salts with a pharmaceutically acceptable acid or base of the PPAR ligand and antioxidant compounds 20 according to the association likewise form an integral part of the invention. Amongst the pharmaceutically acceptable acids there may be mentioned, without implying any limitation, hydrochloric acid, hydrobromic acid, sulphuric acid, phosphonic acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic acid, citric acid, 25 ascorbic acid, oxalic acid, methanesulphonic acid, camphoric acid, etc.. Amongst the pharmaceutically acceptable bases there may be mentioned, without implying any limitation, sodium hydroxide, potassium hydroxide, triethylamine, tert-butylamine, etc.

5a The association to which preference is given in accordance with the invention is rosiglitazone and coenzyme Q1o. Furthermore, the association according to the invention between one or more compounds favouring the lipid and carbohydrate metabolisms and an antioxidant 5 agent has entirely surprising pharmacological properties: the Applicant has discovered that a synergy exists between those two classes of compound allowing a very significant reduction in body fat to be obtained, making it of use in the treatment and/or prevention of obesity and of overweight characterised by a body mass index greater than 25.

-6 In the United States, obesity affects 20 % of men and 25 % of women. Patients having a body mass index (BMI = weight (kg) / height 2

(M

2 )) greater than or equal to 30 are considered to be obese (Int. J. Obes., 1998, 22, 39-47; Obesity Lancet, 1997, 350, 423 426). Obesity (BMI > 30) and overweight (25 < BMI < 30) can have various origins : they 5 may come about following deregulation of food intake, following hormonal disturbance, or following administration of a treatment: treating type II diabetes with sulphonylureas causes patients to gain weight. Similarly, in type I (insulin-dependent) diabetes, insulin therapy is also a cause of weight gain in patients (In Progress in Obesity Research, 8 I1 International Congress on Obesity, 1999, 739-746; Annals of Internal Medicine, 1998, 128 10 165-175). Obesity and overweight are well-established risk factors for cardiovascular diseases: they are associated with a significant increase in the risk of cerebro-vascular accidents and non insulin-dependent diabetes, because they predispose to insulin-resistance, dyslipidaemia and the appearance of macrovascular disorders (nephropathy, retinopathy, angiopathy). 15 Further pathologies are the consequence of obesity or overweight: there may be mentioned, in particular, vesicular calculi, respiratory dysfunction, osteoarthritis, several forms of cancer and, in the case of very severe obesity, premature death (N. Engl. J. Med., 1995, 333, 677-385; JAMA, 1993, 270, 2207-2212). The association according to the invention allows a weight loss to be obtained which, even 20 if moderate, significantly reduces all the risk factors associated with obesity (Int. J. Obes., 1997, 21, 55-9; Int. J. Obes., 1992, 21, S5-9). The association according to the invention will therefore be found to be useful in the treatment and/or prevention of obesity and of overweight characterised by a body mass index greater than 25 and less than 30. 25 The invention accordingly relates to the use of the association comprising one or more PPAR ligands and an antioxidant agent in obtaining pharmaceutical compositions intended for the treatment and/or prevention of obesity and of overweight characterised by a body mass index greater than 25 and less than 30.

-7 In particular, the association according to the invention is of use in the treatment and/or prevention of obesity and of overweight characterised by a body mass index greater than 25 and less than 30 caused by a therapeutic treatment, such as treatment for type I or II diabetes. 5 The invention accordingly relates to the use of an association comprising one or more PPAR ligands and an antioxidant agent in obtaining pharmaceutical compositions intended for the treatment and/or prevention of obesity and of overweight characterised by a body mass index greater than 25 and less than 30 caused by a therapeutic treatment, such as treatment for type I or II diabetes. 10 The invention relates also to pharmaceutical compositions comprising the association between one or more compounds favouring the lipid and carbohydrate metabolisms, more especially one or more PPAR ligands, and an antioxidant agent, as defined hereinbefore, in combination with one or more pharmaceutically acceptable excipients. Among the pharmaceutical compositions according to the invention, there may be 15 mentioned, more especially, those that are suitable for oral, parenteral or nasal administration, tablets or drag6es, sublingual tablets, gelatin capsules, lozenges, suppositories, creams, ointments, dermal gels, etc.. The dosage used varies according to the sex, age and weight of the patient, the administration route, the nature of the therapeutic indication or of any associated 20 treatments and ranges from 0.1 mg to 1 g of each component of the association per 24 hours in one or more administrations. The Examples that follow illustrate the invention but do not limit it in any way. EXAMPLE A: Change in body weight Male C57 Black 6 ob/ob mice from 8 to 12 weeks old were used. The mice are diabetic 25 (type II) and suffer from hyperglycaemia, hypertriglyceridaemia and hyperinsulinaemia. After being placed in quarantine for one week, they were weighed and then randomised as -8 a function of their weight and 6 homogeneous groups (starting weights not significantly different) were formed. After being weighed, the mice were treated with rosiglitazone (antidiabetic agent) on its own or in association with coenzyme Qi. The compounds were injected by the intraperitoneal route once a day for 14 days in a solution of DMSO 5 % / 5 Solutol 15 % / qsp H 2 0 heated at 65*C to ensure good dissolution. In addition, the solution was pre-heated before injection. The mice were weighed every day and the weight obtained after 14 days of treatment was recorded. Treatment with rosiglitazone alone results in an increase in the weight of the mice greater than or equal to 5 grams, corresponding to about 10 % more than their initial weight. The 10 association rosiglitazone+coenzyme Q o allows that weight gain to be reversed by at least 50 % and demonstrates the effectiveness of the association in reducing body weight. EXAMPLE B : Pharmaceutical composition 100 tablets each containing 30 mg of rosiglitazone and 10 mg of coenzyme Qio R osiglitazone ......................................................................................................... 3 g 15 C oenzym e Q io .................................................................................................. 1 g W heat starch ........................................................................................................

20 g M aize starch ........................................................................................................ 20 g L actose ................................................................................................................ 30 g M agnesium stearate...............................................................................................2 g 20 S ilica ......................................................................................................................

1 g H ydroxypropylcellulose........................................................................................2 g 9 The Examples that follow demonstrate synergy between the two classes of compounds of the invention. Two additional examples of association of 1) a mixed PPARa/y ligand or a 5 PPARa and a PPARy ligand with 2) an antioxidant agent demonstrate the surprising effect of the associations according to the invention: Common protocol to the two flowing examples: Male C57 Black 6 ob/ob mice from 8 to 12 weeks old were used. After being 10 placed in quarantine for one week, they were weighed and then randomised as a function of their weight and 6 homogeneous groups (starting weights not significantly different) were formed. After being weighed, the various compounds under test were injected by the intraperitoneal route once a day for 7 days. The compounds were injected in a solution of DMSO 5 % / Solutol 15 % / qsp H 2 0 15 heated at 65 0 C to ensure good dissolution. In addition, the solution was pre heated before injection. The mice were weighed every day and the weight obtained after 7 days of treatment was recorded. EXAMPLE 20 A reduction of 30 % is observed in the weight gain of mice treated with the association comprising 2-ethoxy-3-{4-[2-(6-[(hydroxyimino)(phenyl)methyl]-2-oxo 1,3-benzothiazol-3(2H)-yl)ethoxy]phenyl}propanoic acid which is a PPARa/y ligand and coenzyme Q10, whereas coenzyme Q10 administered on its own does 25 not reduce the weight gain and 2-ethoxy-3-{4-[2-(6 [(hydroxyimino)(phenyl)methyl]-2-oxo-1,3-benzothiazol-3(2H)-yl)ethoxy]phenyl} propanoic acid administered on its own reduces the weight gain by only 10 %. EXAMPLE 2 30 The results obtained with the association of fenofibrate which is a PPARa ligand + rosiglitazone which is a PPARy ligand + coenzyme Q10 are indicated below and are expressed as the percentage weight change with respect to the control (corresponding to mice treated with the injection solvent for 7 days): 10 I -I 10.0 -Control - PPARy 10 mg/kg 7.5- F-1 M CoQ 10 mg/kg MPPARy/CoQ 5.0 || PPARy/PPARa/CoQ 2.5 0.01 Therefore, the results obtained from examples 1 and 2 clearly show: - that the association enables the weight of obese mice to be reduced 5 significantly, - that there exists a synergy between the 2 or 3 components of the association, the weight loss found being much greater in the case of the association than in the case of each component administered on its own. 10 The following data correspond to quantitative data of example A in the description of the present Australian Patent Application. We observe that the weight of obese mice is reduced of 2.66g between mice treated with PPARy (6.52g +/-0.91) and mice treated with PPARy + coenzyme Q1o (3.86g +/-0.36). Thus in view of these quantitative data, the weight loss is less or equal to 2.5 grams in accordance with 15 the previous argument. Control : 2.86 +/- 0.38 (n=8) PPARy (Rosiglitazone)10 mg/kg 6.52 +/-0.91 (n= 7) CoQ 1 O mg/kg 2.98 +/- 0.18 (n=8) 20 PPARy (Rosiglitazone)/CoQ 1 o: 3.86 +/ 0.36 (n=6) PPARy (Rosiglitazone)/PPARa/CoQ : 0.36 +/-0.97 (n=6) 11 * 8 Control 6 PPARy 10 mg/kg 5 ' CoQ 10 mg/kg M PPARy/CoQ 4 ||| PPARy/PPARa/CoQ 100 The associations of a mixed PPARa/y ligand or a PPARa ligand and a PPARy ligand combined with an antioxidant disclosed in the invention have surprising properties in the treatment or prevention of obesity. Indeed, the associations 15 enable the weight to be reduced significantly and allow a synergy between the 2 or 3 components of the association since the weight loss found being much greater in the case of the association than in the case of each component administered on its own. These surprising properties of these associations are confirmed and supported by the above comparative trials. 20 ADDITIONAL EXAMPLES Two additional examples with regard to Australian Patent application 2003299772 of association of 1) a mixed PPARa/y ligand or a PPARa and a PPARy ligand 25 with 2) an antioxidant agent demonstrate the surprising effect of the associations according to the invention. Common protocol to the two flowing examples: 30 Male C57 Black 6 ob/ob mice from 8 to 12 weeks old were used. After being placed in quarantine for one week, they were weighed and then randomised as a function of their weight and 6 homogeneous groups (starting weights not significantly different) were formed. After being weighed, the various compounds 12 under test were injected by the intraperitoneal route once a day for 7 days. The compounds were injected in a solution of DMSO 5 % / Solutol 15 % I qsp H20 heated at 65 0 C to ensure good dissolution. In addition, the solution was pre heated before injection. The mice were weighed every day and the weight 5 obtained after 7 days of treatment was recorded. EXAMPLE 3 A reduction of 30 % is observed in the weight gain of mice treated with the 10 association comprising 2-ethoxy-3-{4-[2-(6-[(hydroxyimino)(phenyl)methyl]-2-oxo 1,3-benzothiazol-3(2H)-yl)ethoxy]phenyl}propanoic acid which is a PPARa/y ligand and coenzyme Q 1 0 , whereas coenzyme Q10 administered on its own does not reduce the weight gain and 2-ethoxy-3-{4-[2-(6 [(hydroxyimino)(phenyl)methyl]-2-oxo-1,3-benzothiazol-3(2H)-yl)ethoxy]phenyl} 15 propanoic acid administered on its own reduces the weight gain by only 10 %. EXAMPLE 4 The results obtained with the association of fenofibrate which is a PPARa ligand + 20 rosiglitazone which is a PPARy ligand + coenzyme Q 1 0 are indicated below and are expressed as the percentage weight change with respect to the control (corresponding to mice treated with the injection solvent for 7 days): I I 10.0 1 0 .1- C o n tro l 7.5- MPPARy 10 mg/kg j7-l CoQ 10 mg/kg PPARy/CoQ .:A5.0 [il PPARy/PPARax/CoQ 2.5 0.011 l] 13 Therefore, the results obtained from examples 1 and 2 clearly show: - that the association enables the weight of obese mice to be reduced significantly, 5 - that there exists a synergy between the 2 or 3 components of the association, the weight loss found being much greater in the case of the association than in the case of each component administered on its own. The following data correspond to quantitative data of example A in the description 10 of the Indian patent applicationl381/DELNP/2005. We observe that the weight of obese mice is reduced of 2.66g between mice treated with PPARy (6.52g +/-0.91) and mice treated with PPARy + coenzyme Q 1 0 (3.86g +/-0.36). Thus in view of these quantitative data, the weight loss is less or equal to 2.5 grams in accordance with the previous argument. 15 Control : 2.86 +/- 0.38 (n=8) PPARy (Rosiglitazone)10 mg/kg 6.52 +/-0.91 (n= 7) CoQ 1 o mg/kg 2.98 +/- 0.18 (n=8) PPARy (Rosiglitazone)/CoQ1o: 3.86 +/ 0.36 (n=6) 20 PPARy (Rosiglitazone)/PPARa/CoQ : 0.36 +/-0.97 (n=6) 25 8- Control MPPARy 10 mg/kg i 6CoQ 10 mg/kg & PPARy/CoQ .c4 .P [l PPARy/PPARa/CoQ 30 2 0 14 Once again the associations of a mixed PPARa/y ligand or a PPARa ligand and a PPARy ligand combined with an antioxidant disclosed in the invention have surprising properties in the treatment or prevention of obesity. Indeed, the 5 associations enable the weight to be reduced significantly and allow a synergy between the 2 or 3 components of the association since the weight loss found being much greater in the case of the association than in the case of each component administered on its own. These surprising properties of these associations are confirmed and supported by the above comparative trials. 10 Comprises/comprising and grammatical variations thereof when used in this specification are to be taken to specify the presence of stated features, integers, steps or components or groups thereof, but do +not preclude the presence or addition of one or more other features, integers, steps, components or groups 15 thereof.

Claims (12)

1. Composition comprising 1) an antioxidant agent and 2) a mixed PPAR ligand for the a and y receptor sub-types, or a selective PPAR ligand for the a receptor sub-type and a selective PPAR ligand for the y receptor sub-type 5 optionally as a pharmaceutical composition being in combination with one or more pharmaceutically acceptable excipients.

2. Composition according to claim 1, wherein the PPAR ligand is a mixed ligand for the a and y receptor sub-types.

3. Composition according to claim 1, comprising a selective ligand for the a 10 receptor sub-type and a selective ligand for the y receptor sub-type.

4. Composition according to claim 1, wherein the selective PPAR ligand for the y receptor sub-type is rosiglitazone or an addition salt thereof with a pharmaceutically acceptable acid or base.

5. Composition according to claim 1, wherein the antioxidant agent is 15 ubiquinone or coenzyme Q 10 .

6. Composition, which is rosiglitazone and coenzyme Q 10 optionally as a pharmaceutical composition being in combination with one or more pharmaceutically acceptable excipients.

7. Composition which is substantially as hereinbefore described with 20 reference to Example B.

8. Use of a composition according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment and/or prevention of obesity.

9. Use of a composition according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of obesity caused by a 25 therapeutic treatment. 16

10. Use of a composition according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of obesity caused by treatment for type I or 11 diabetes.

11. Use of a composition according to any one of claims 1 to 7 in the 5 manufacture of a medicament for the treatment and/or prevention of being overweight characterised by a body mass index greater than 25 and less than

30. 12. Use of a composition according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of being overweight 10 characterised by a body mass index greater than 25 and less than 30 caused by a therapeutic treatment. 13. Use of a composition according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment of being overweight characterised by a body mass index greater than 25 and less than 30 caused by 15 treatment for type I or il diabetes. 14. A method for the treatment and/or prevention of obesity in a subject, which method comprises administering to the subject a therapeutic amount of a pharmaceutical composition according to any one of claims 1 to 7. 15. A method for the treatment and/or prevention of obesity in a subject 20 caused by a therapeutic treatment, which method comprises administering to the subject a therapeutic amount of a pharmaceutical composition according to any one of claims 1 to 7. 16. A method for the treatment and/or prevention of obesity in a subject caused by a treatment for type I or 11 diabetes, which method comprises 25 administering to the subject a therapeutic amount of a pharmaceutical composition according to any one of claims 1 to 7. 17 17. A method for the treatment and/or prevention of being overweight characterised by a body mass index greater than 25 and less than 30, which method comprises administering to an overweight subject a therapeutic amount of a pharmaceutical composition according to any one of claims 1 to 7. 5 18. A method for the treatment and/or prevention of being overweight characterised by a body mass index greater than 25 and less than 30, caused by a therapeutic treatment, which method comprises administering to an overweight subject a therapeutic amount of a pharmaceutical composition according to any one of claims 1 to 7. 10 19. A method for the treatment and/or prevention of being overweight characterised by a body mass index greater than 25 and less than 30, caused by a treatment for type I or 11 diabetes, which method comprises administering to an overweight subject a therapeutic amount of a pharmaceutical composition according to any one of claims 1 to 7. 15 LES LABORATOIRES SERVIER & CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE 20 WATERMARK PATENT & TRADE MARK ATTORNEYS P25404AU00